Etiologic Risk Group (see link below): */Disease Information (sort of like the Intro to your Mini Research Write up):
Tuberculosis is commonly known as a respiratory infection caused by Mycobacterium tuberculosis and can become fatal if not treated. It can be spread through the air by people who are already infected. The symptoms of tuberculosis include chest pains, coughing up blood or phlegm, and a bad cough that can last 3 weeks or longer.
Link to TDR Targets page (if present):
N/A
Link to Gene Database page (NCBI, EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.)
Essentiality of this protein:
This protein is essential to the ESX-1 secretion apparatus, which provides the principal virulence factors ESAT-6 and CFP-10 Is it a monomer or multimer as biological unit? (make prediction athttp://www.ebi.ac.uk/msd-srv/prot_int/pistart.html): Complex of proteins?:
No
Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism):
ATP Protein + L-histidine = ADP + Protein N(tau)-phospho-L-histidine
-- Show screenshot of BRENDA enzyme mechanism schematic Enzyme Assay information (spectrophotometric, coupled assay ?, reagents):
Supernatant fluid was filtered and then run through an enzyme-linked immunosorbent assay (ELISA)
-- link to Sigma (or other company) page for assay (see Sigma links below) -- -or link (or citation) to paper that contains assay information
-- links to assay reagents (substrates) pages. --- List cost and quantity of substrate reagents, supplier, and catalog #
Structure (PDB or Homology model) -- PDB # or closest PDB entry if using homology model: -- For Homology Model option: ---- Show pairwise alignment of your BLASTP search in NCBI against the PDB ---- Query Coverage: ---- Max % Identities: ---- % Positives ---- Chain used for homology:
PDB #: P9WJE0 (no PDB entries available for this one or homologs)
Current Inhibitors: Expression Information (has it been expressed in bacterial cells): Purification Method: Purified with the use of nickel agarose Image of protein (PyMol with features delineated and shown separately):
*Amino Acid Sequence (paste as text only - not as screenshot or as 'code'):
*CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only): *GC% Content for gene (codon optimized):
Do Not Need this info for Spring (but still copy these lines to your Target page for now) Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers): (link to DNA Works output text file - that should be saved in your Google Docs folder after you did the primer design protocol) -- Ask a mentor, Dr. B, or a fellow researcher -how to link a GDocs file if you are not sure how to.
Primer design results for 'tail' primers (this is just 2 sequences): **
EspA
*NCBI Gene # or RefSeq#:
885377
*Protein ID (NP or XP #) or Wolbachia#:
NP_218133.1
*Organism (including strain):
Mycobacterium tuberculosis
Etiologic Risk Group (see link below):
*/Disease Information (sort of like the Intro to your Mini Research Write up):
Tuberculosis is commonly known as a respiratory infection caused by Mycobacterium tuberculosis and can become fatal if not treated. It can be spread through the air by people who are already infected. The symptoms of tuberculosis include chest pains, coughing up blood or phlegm, and a bad cough that can last 3 weeks or longer.
Link to TDR Targets page (if present):
N/A
Link to Gene Database page (NCBI, EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.)
https://www.ncbi.nlm.nih.gov/gene/885377
Essentiality of this protein:
This protein is essential to the ESX-1 secretion apparatus, which provides the principal virulence factors ESAT-6 and CFP-10
Is it a monomer or multimer as biological unit? (make prediction at http://www.ebi.ac.uk/msd-srv/prot_int/pistart.html):
Complex of proteins?:
No
Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism):
http://jb.asm.org/content/195/24/5421.full
*EC#:
2.7.13.3
Link to BRENDA EC# page:
http://www.brenda-enzymes.org/enzyme.php?ecno=2.7.13.3&Suchword=&reference=&UniProtAcc=&organism%5B%5D=Mycobacterium+tuberculosis+H37Rv
ATP Protein + L-histidine = ADP + Protein N(tau)-phospho-L-histidine
-- Show screenshot of BRENDA enzyme mechanism schematic
Enzyme Assay information (spectrophotometric, coupled assay ?, reagents):
Supernatant fluid was filtered and then run through an enzyme-linked immunosorbent assay (ELISA)
-- link to Sigma (or other company) page for assay (see Sigma links below)
-- -or link (or citation) to paper that contains assay information
http://jb.asm.org/content/195/24/5421.full
-- links to assay reagents (substrates) pages.
--- List cost and quantity of substrate reagents, supplier, and catalog #
Structure (PDB or Homology model)
-- PDB # or closest PDB entry if using homology model:
-- For Homology Model option:
---- Show pairwise alignment of your BLASTP search in NCBI against the PDB
---- Query Coverage:
---- Max % Identities:
---- % Positives
---- Chain used for homology:
PDB #: P9WJE0 (no PDB entries available for this one or homologs)
Current Inhibitors:
Expression Information (has it been expressed in bacterial cells):
Purification Method:
Purified with the use of nickel agarose
Image of protein (PyMol with features delineated and shown separately):
*Amino Acid Sequence (paste as text only - not as screenshot or as 'code'):
10 20 30 40 50 MSRAFIIDPT ISAIDGLYDL LGIGIPNQGG ILYSSLEYFE KALEELAAAF 60 70 80 90 100 PGDGWLGSAA DKYAGKNRNH VNFFQELADL DRQLISLIHD QANAVQTTRD 110 120 130 140 150 ILEGAKKGLE FVRPVAVDLT YIPVVGHALS AAFQAPFCAG AMAVVGGALA 160 170 180 190 200 YLVVKTLINA TQLLKLLAKL AELVAAAIAD IISDVADIIK GTLGEVWEFI 210 220 230 240 250 TNALNGLKEL WDKLTGWVTG LFSRGWSNLE SFFAGVPGLT GATSGLSQVT 260 270 280 290 300 GLFGAAGLSA SSGLAHADSL ASSASLPALA GIGGGSGFGG LPSLAQVHAA 310 320 330 340 350 STRQALRPRA DGPVGAAAEQ VGGQSQLVSA QGSQGMGGPV GMGGMHPSSG 360 370 380 390 ASKGTTTKKY SEGAAAGTED AERAPVEADA GGGQKVLVRN VV*length of your protein in Amino Acids
392
Molecular Weight of your protein in kiloDaltons using the Expasy ProtParam website
39.89 kDa
Molar Extinction coefficient of your protein at 280 nm wavelength:
0.951 L/mol cm
TMpred graph Image (http://www.ch.embnet.org/software/TMPRED_form.html). Input your amino acid sequence to it.
*CDS Gene Sequence (paste as text only):
*GC% Content for gene:
63.4%
*CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only):
*GC% Content for gene (codon optimized):
Do Not Need this info for Spring (but still copy these lines to your Target page for now)
Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers):
(link to DNA Works output text file - that should be saved in your Google Docs folder after you did the primer design protocol)
-- Ask a mentor, Dr. B, or a fellow researcher -how to link a GDocs file if you are not sure how to.
Primer design results for 'tail' primers (this is just 2 sequences):
**